Open AccessSequence of pig lens aldose reductase and electrospray mass spectrometry of non‐covalent and covalent complexes
Author(s) -
JAQUINOD Michel,
POTIER Noelle,
KLARSKOV Klaus,
REYMANN JeanMarc,
SOROKINE Odile,
KIEFFER Sylvie,
BARTH Patrick,
ANDRIANTOMANGA Verotiana,
BIELLMANN JeanFrançois,
DORSSELAER Alain
Publication year - 1993
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1993.tb18445.x
Subject(s) - covalent bond , aldose reductase , chemistry , mass spectrometry , sequence (biology) , electrospray ionization , lens (geology) , electrospray , biochemistry , chromatography , enzyme , biology , organic chemistry , paleontology
The complete sequence of pig lens aldose reductase (EC 1.1.1.21), a member of the nicotinamide coenzyme‐dependent aldo‐keto reductase super family, was determined by the combined use of data obtained from Edman degradation, fast‐atom‐bombardment mass spectrometry and electrospray mass spectrometry. The N‐terminal residue of human and pig aldose reductase was shown to be acetylated. The assignment of a disulfide bridge (Cys298–Cys303) was obtained by mass spectrometry. Electrospray mass spectrometry has been used for molecular mass measurement of human muscle (35758±7Da) and pig lens (35778±3Da) aldose reductase; using mild ionization conditions, it has also been used to study the reversible interaction involved in a non‐covalent complex with NADP + (36527±4Da). An alkylating analog of NADP + (3‐chloroacetylpyridine–adenine dinucleotide phosphate) was used as an irreversible inhibitor to investigate the NADP binding site and the mass of the covalent complex was measured (36521±3Da).